使用核孔蛋白偶联磁珠研究核膜上的核孔复合体组装
Use of Nucleoporin-Conjugated Beads to Study the Nuclear Pore Complex Assembly on the Nuclear Membrane.
作者信息
Bilir Şükriye, Hiraoka Yasushi, Haraguchi Tokuko
机构信息
Graduate School of Frontier Biosciences, Osaka University, Suita, Japan.
Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, Türkiye.
出版信息
Methods Mol Biol. 2025;2958:169-181. doi: 10.1007/978-1-0716-4714-1_12.
Reassembly of the nuclear pore complex (NPC) at the end of mitosis is a key event for establishing a functional nuclear envelope. However, the role of each NPC component (Nup) in NPC reassembly remained unclear. Here, we describe a method using artificial beads conjugated with an anti-GFP antibody. In this method, such beads are introduced into living cells expressing the Nup of interest fused with GFP to tether the specific Nup around the beads. By examining the Nup-conjugated beads in mitotic telophase cells, it is possible to investigate the role of the specific Nup of interest in NPC formation on the nuclear membrane.
有丝分裂末期核孔复合体（NPC）的重新组装是建立功能性核膜的关键事件。然而，每个NPC组分（核孔蛋白）在NPC重新组装中的作用仍不清楚。在此，我们描述了一种使用与抗GFP抗体偶联的人工珠子的方法。在这种方法中，将此类珠子引入表达与GFP融合的目标核孔蛋白的活细胞中，以便将特定的核孔蛋白束缚在珠子周围。通过检查有丝分裂末期细胞中与核孔蛋白偶联的珠子，有可能研究目标特定核孔蛋白在核膜上NPC形成中的作用。
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